PROJECT SUMMARY There is an important unmet need in glaucoma to develop next-generation functional imaging modalities. The advent of high resolution retinal imaging technologies, specifically, optical coherence tomography and adaptive optics scanning laser ophthalmoscopy, has enabled evaluation of retinal anatomical features at a cellular level in living human eyes. However, assessment of anatomy only provides limited information about the health and function of the retinal tissue. There remains a pressing need for molecular and metabolic endpoints in evaluating glaucoma, where structural changes may take many years to appear, often associated with significant vision loss. Currently available techniques for assessment of retinal blood flow, oxygen saturation, and mitochondrial function in humans have limited depth discrimination and require separate instruments, hampering accurate and comprehensive quantification of retinal ganglion cell physiology. Thus, we propose to develop and validate an innovative depth-resolved retinal metabolic imaging system, scanning protocols and analysis algorithms to non-invasively, quantitatively and simultaneously assess retinal oxygen and mitochondrial metabolism in glaucoma, and in response to candidate neuroprotective or regenerative therapies, both in animal models and in human subjects. We will perform pilot studies to demonstrate and validate the technology's capability to provide metabolic measures relevant to vision restorative therapies in animals and humans. The availability of novel retinal metabolic measures will for the first time introduce a physiologically-based outcome measure for disease characterization and also for candidate therapies relevant to regenerative ophthalmology and vision restoration. In addition, the proposed research will have a significant impact on advancing: 1) knowledge of retinal metabolic dysfunction in animal models of human glaucoma and other optic neuropathies, 2) translation of candidate regenerative therapies that improve retinal metabolic function in pre-clinical studies, and 3) clinical assessment and evaluation of available and emerging therapies for restoring vision in glaucoma, as well as in other degenerative retina/optic nerve diseases.